title: Generation of a large number of functional dendritic cells from human monocytes expanded by forced expression of cMYC plus BMI1.
abstract: Anticancer vaccination therapies with monocyte-derived dendritic cells (DC) are widely conducted. A large number of primary monocytes (approximately 10(8) cells) are needed to generate the number of DC required to achieve an effect upon vaccination, and monocytes are usually purified from peripheral blood mononuclear cells obtained by apheresis procedure, which is somehow invasive for cancer patients. As a means to facilitate the generation of DC for therapeutic use, we herein report a method to amplify human monocytes. We found that lentivirus-mediated transduction of cMYC along with BMI1 induced proliferation of CD14(+) monocytes derived from 9 out of 12 blood donors, and we named the monocyte-derived proliferating cells CD14-ML. Their proliferation continued for 3-5 weeks in the presence of M-CSF and GM-CSF, resulting in 20-1000-fold amplification. Importantly, the expanded CD14-ML differentiated into fully functional DC (CD14-ML-DC) upon the addition of IL-4 to the culture. We successfully stimulated autologous CD8(+) T cells with CD14-ML-DC pulsed with cytomegalovirus peptide or MART-1 peptide to generate antigen-specific CTL lines. This is the first report describing the method for in vitro expansion of human peripheral blood monocytes.
authors.affiliation: Department of Immunogenetics, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan; CREST, Japan Science and Technology Agency, Kawaguchi, Japan.
authors.forename: Miwa
authors.initials: M
authors.lastname: Haruta
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Tomita
authors.forename: Yuya
authors.initials: Y
authors.lastname: Imamura
authors.forename: Keiko
authors.initials: K
authors.lastname: Matsumura
authors.forename: Tokunori
authors.initials: T
authors.lastname: Ikeda
authors.forename: Koutaro
authors.initials: K
authors.lastname: Takamatsu
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Nishimura
authors.forename: Satoru
authors.initials: S
authors.lastname: Senju
mesh_terms: D017951:Antigen Presentation / Q000276:immunology; D019496:Cancer Vaccines / Q000276:immunology; D002454:Cell Differentiation; D002460:Cell Line; D003587:Cytomegalovirus / Q000276:immunology; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000235:genetics / Q000378:metabolism; D058965:MART-1 Antigen / Q000276:immunology; D009000:Monocytes / Q000166:cytology* / Q000276:immunology / Q000378:metabolism*; D022423:Myeloid Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D063150:Polycomb Repressive Complex 1 / Q000235:genetics*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics*; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism
chemical_list: C069077:BMI1 protein, human; D019496:Cancer Vaccines; D018950:Lipopolysaccharide Receptors; D058965:MART-1 Antigen; D016271:Proto-Oncogene Proteins c-myc; D063150:Polycomb Repressive Complex 1